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刊物信息

期刊名称:药物分析杂志
主管单位:中国科学技术协会
主办单位:中国药学会
承办:中国食品药品检定研究院
主编:金少鸿
地址:北京天坛西里2号
邮政编码:100050
电话:010-67012819,67058427
电子邮箱:ywfx@nicpbp.org.cn
国际标准刊号:ISSN 0254-1793
国内统一刊号:CN 11-2224/R
邮发代号:2-237
 

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抗PD-L1单抗的质量控制研究

Study on quality control of anti-PD-L1 monoclonal antibody

作者(英文):
分类号:R917
出版年·卷·期(页码):2019,39 (1):13-22
DOI: 10.16155/j.0254-1793.2017.01.01
-----摘要:-------------------------------------------------------------------------------------------

目的:研究建立抗程序性死亡受体配体1(programmed cell death protein ligand 1,PD-L1)单抗的质量控制方法。方法:利用成像毛细管等电聚焦电泳(imaging capillary isoelectric focusing electrophoresis,iCIEF)、肽图对抗PD-L1单抗进行鉴别;利用还原/非还原十二烷基磺酸钠毛细管电泳(capillary electrophoresis-sodium dodecyl sulfonate,CE-SDS)、分子排阻高效液相色谱(size exclusion-high performance liquid chromatography,SEC-HPLC)、离子交换高效液相色谱(ion exchange-high performance liquid chromatography,IEC-HPLC)对抗PD-L1单抗的纯度进行控制;利用细胞结合抑制法和转基因细胞法测定抗PD-L1单抗的活性;利用光阻法和液流成像分析法对不溶性微粒进行检测和分析。结果:抗PD-L1单抗的iCIEF检测结果具有特征性主峰,肽图检测具有相应特征峰,起到很好的鉴别作用。还原CE-SDS的轻链与重链峰面积之和的百分比为(98.37±0.21)%,非还原CE-SDS主峰峰面积百分比为(96.33±0.15)%;SEC-HPLC单体的峰面积百分比为(99.43±0.02)%;IEC-HPLC酸性区峰面积百分比、主峰峰面积百分比、碱性区峰面积百分比分别为(13.63±0.40)%、(80.00±0.36)%、(6.37±0.15)%。细胞结合抑制法与转基因细胞法的活性检测均呈现剂量反应曲线,反映抗PD-L1单抗剂量依赖性地阻断程序性死亡受体1(programmed cell death protein 1,PD-1)与PD-L1的结合及后续的生物学效应。光阻法检测≥10 μm及≥25 μm的不溶性微粒数分别为(1.33±0.58)粒·mL-1和(0.00±0.00)粒·mL-1;液流成像分析法检测2~10 μm、≥10 μm及≥25 μm的微粒数分别为(1 243.67±242)粒·mL-1、(45.67±16.07)粒·mL-1和(10.00±5.00)粒·mL-1,其中,蛋白聚体占87.45%,非蛋白聚体占12.55%。结论:研究建立了抗PD-L1单抗的多种质控方法,为国内抗PD-L1单抗的质量检测提供了参考依据。  

-----英文摘要:---------------------------------------------------------------------------------------

Objective:To investigate and establish quality control methods for anti-programmed cell death protein ligand 1(PD-L1) monoclonal antibody(mAb). Methods:Imaging capillary isoelectric focusing electrophoresis(iCIEF) and peptide mapping were used for identification of the anti-PD-L1 mAb. The purity was measured using mothods including reduced/non-reduced capillary electrophoresis-sodium dodecyl sulfonate(CE-SDS),size exclusion-high performance liquid chromatography (SEC-HPLC) and ion exchange-high performance liquid chromatography(IEC-HPLC). The activity of anti-PD-L1 mAb was determined by cell binding inhibition method and transgenic cell method,and the sub-visible particles were detected and analyzed by light obscuration and flow imaging method. Results:The iCIEF and peptide mapping data showed the characteristic peaks of anti-PD-L1 mAb,which played a promising role in product identification. The area percentage of the sum of both light chain and heavy chain was (98.37±0.21)%,as detected by reduced CE-SDS assay;the area percentage of the main peak detected by non-reduced CE-SDS was (96.33±0.15)%. The area percentage of monomer peak detected by SEC-HPLC was (99.43±0.02)%. The area percentage of the acid peak,the main peak and the basic peak were (13.63±0.40)%,(80.00±0.36)% and (6.37±0.15)%,respectively. In bio-specific activity assays,both cell binding inhibition method and transgenic cell method assays showed dose-response curves,reflecting dose-dependent blockade of the binding between PD-L1 and programmed cell death protein 1(PD-1),as well as the subsequent anti-PD-L1 mAb-induced biological effect. The detected amount of sub-visible particles with diameters of ≥ 10 μm, ≥ 25 μm by light obscuration were (1.33±0.58) particle·mL-1 and (0.00±0.00) particle·mL-1,respectively;the detected amount of sub-visible particles with diameters of 2-10 μm, ≥ 10 μm and ≥ 25 μm by flow imaging method were (1 243.67±242) particle·mL-1,(45.67±16.07) particle·mL-1 and (10.00±5.00) particle·mL-1,respectively,in which 87.45% were protein aggregates while 12.55% were non-protein particles. Conclusion:A series of representive quality control methods for anti-PD-L1 mAb have been established,which can provide references for the quality control of anti-PD-L1 mAb in China. 

-----参考文献:---------------------------------------------------------------------------------------

[1] PARDOLL DM.The blockade of immune checkpoints in cancer immunotherapy[J].Nat Rev Cancer,2012,12(4):252
[2] KEIR ME,LIANG SC,INDIRA G,et al.Tissue expression of PD-L1 mediates peripheral T cell tolerance[J].J Exp Med,2006,203(4):883
[3] FRANCISCO LM,SALINAS VH,BROWN KE,et al.PD-L1 regulates the development,maintenance,and function of induced regulatory T cells[J].Clin Immunol,2009,131(13):S41
[4] OKAZAKI T,HONJO T.PD-1 and PD-1 ligands:from discovery to clinical application[J].Int Immunol,2007,19(7):813
[5] IWAI Y,ISHIDA M,TANAKA Y,et al.Involvement of PD-L1 on tumor cells in the escape from host immune system and tumor immunotherapy by PD-L1 blockade[J].Proc Natl Acad Sci USA,2002,99(19):12293
[6] BLANK C,MACKENSEN A.Contribution of the PD-L1/PD-1 pathway to T-cell exhaustion:an update on implications for chronic infections and tumor evasion[J].Cancer Immunol Immunother,2007,56(5):739
[7] GAO Q,WANG XY,QIU SJ,et al.Overexpression of PD-L1 significantly associates with tumor aggressiveness and postoperative recurrence in human hepatocellular carcinoma[J].Clin Cancer Res,2009,15(3):971
[8] MU CY,HUANG JA,CHEN Y,et al.High expression of PD-L1 in lung cancer may contribute to poor prognosis and tumor cells immune escape through suppressing tumor infiltrating dendritic cells maturation[J].Med Oncol,2011,28(3):682
[9] BLANK C,KUBALL J,VOELKL S,et al.Blockade of PD-L1(B7-H1) augments human tumor-specific T cell responses in vitro[J].Int J Cancer,2010,119(2):317
[10] TOPALIAN SL,DRAKE CG,PARDOLL DM.Targeting the PD-1/B7-H1(PD-L1) pathway to activate anti-tumor immunity[J].Curr Opin Immunol,2012,24(2):207
[11] CHEN L,HAN X.Anti-PD-1/PD-L1 therapy of human cancer:past,present,and future[J].J Clin Invest,2015,125(9):3384
[12] SWAIKA A,HAMMOND WA,JOSEPH RW.Current state of anti-PD-L1 and anti-PD-1 agents in cancer therapy[J].Mol Immunol,2015,67(2):4
[13] TENG F,MENG X,KONG L,et al.Progress and challenges of predictive biomarkers of anti PD-1/PD-L1 immunotherapy:a systematic review[J].Cancer Lett,2018,414:166
[14] HERBST RS,SORIA JC,KOWANETZ M,et al.Predictive correlates of response to the anti-PD-L1 antibody MPDL3280A in cancer patients[J].Nature,2014,515(7528):563
[15] POWLES T,EDER JP,FINE GD,et al.MPDL3280A (anti-PD-L1) treatment leads to clinical activity in metastatic bladder cancer[J].Nature,2014,515(7528):558
[16] KOTSAKIS A,GEORGOULIAS V.Avelumab,an anti-PD-L1 monoclonal antibody,shows activity in various tumour types[J].Lancet Oncol,2017,18(5):556
[17] RAMOS-ESQUIVEL A,van der LAAT A,ROJAS-VIGOTT R,et al.Anti-PD-1/anti-PD-L1 immunotherapy versus docetaxel for previously treated advanced non-small cell lung cancer:a systematic review and meta-analysis of randomised clinical trials[J].ESMO Open,2017,2(3):e000236
[18] VALECHA GK,VENNEPUREDDY A,IBRAHIM U,et al.Anti-PD-1/PD-L1 antibodies in non-small cell lung cancer:the era of immunotherapy[J].Expert Rev Anticancer Ther,2017,17(1):47
[19] ADCOCK JL,BARROW CJ,BARNETT NW,et al.Chemiluminescence and electrochemiluminescence detection of controlled drugs[J].Drug Test Anal,2011,3(3):145
[20] WANG L,YU C,YANG Y,et al.Development of a robust reporter gene assay to measure the bioactivity of anti-PD-1/anti-PD-L1 therapeutic antibodies[J].J Pharm Biomed Anal,2017,145:447
[21] ANDERSON CL,WANG Y,RUSTANDI RR.Applications of imaged capillary isoelectric focussing technique in development of biopharmaceutical glycoprotein-based products[J].Electrophoresis,2012,33(11):1538

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